Enhanced Wnt sensitivity of CML in comparison with normal stem/progenitor cells. (A) Normal and CML CD34⁺ cells were cultured in recombinant Wnt3A (200 ng/mL), and cells were harvested after 4 h for Western blotting for P-LRP6, LRP6, β-catenin, and GAPDH. (B) Results of densitometry analysis are shown. (C) Normal (CB) and CML CD34⁺ cells (n = 3) cultured in recombinant Wnt3A (200 ng/mL) were plated in methylcellulose assay, following which colonies were counted after 2 weeks of culture. (D) Results of qPCR analysis was performed for FZD genes in normal and CML CD34⁺ cells. CML CD34⁺ cells (n = 3) transfected with control siRNA, FZD1 siRNA, FZD4 siRNA, and FZD5 siRNA were cultured with or without NIL in the absence (E) or presence of MSC (F), for 4 days, following which CFC frequency was analyzed in methylcellulose progenitor assays. Error bars represent mean ± SEM. *P < .05; **P < .01. CB, cord blood.